Congenital Adrenal Hyperplasia (CAH) is a disease associated with mutations in the cytochrome p450 (CYP) 21A2 (CYP21A2) gene, which leads to impaired function and reduced levels of the enzyme 21-hydroxylase (21OHD) that can be classified into: severe salt-wasting (SWCAH), simple virilizing (SVCAH), and non-classical (NCAH) 1-4 . Clinical research is very important in identifying genetic mutations implicated in this disease. Long Range PCR is used to amplify CYP21A2, allowing differentiation between the active gene and non-functioning pseudogene, followed by targeted amplification of all exons within the active gene. This approach allows for 10-20x enrichment of CYP21A2 reads, and mutations detected are exclusively from the functional CYP21A2 locus. NEXTFLEX ® Congenital Adrenal Hyperplasia Amplicon Panel NEWBORN SYNDROMES & INFERTILITY Gene Content CYP21A2 exons Low Sample Requirement > 20 ng DNA from fresh or frozen samples Coverage Uniformity 100% at 0.2x mean coverage On-target Reads > 92% High Sample Indexing Up to 384 samples with 100x coverage on a single Illumina® 2x150 MiSeq® lane for germline mutations Efficient Workflow < 3 hrs hands-on time PERFORMANCE INNOVACIÓN TECNOLÓGICA PARA LABOTORIO
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Congenital Adrenal Hyperplasia (CAH) is a disease associated with mutations in the cytochrome p450 (CYP) 21A2 (CYP21A2) gene, which leads to impaired function and reduced levels of the enzyme 21-hydroxylase (21OHD) that can be classified into: severe salt-wasting (SWCAH), simple virilizing (SVCAH), and non-classical (NCAH)1-4. Clinical research is very important in identifying genetic mutations implicated in this disease.
Long Range PCR is used to amplify CYP21A2, allowing differentiation between the active gene and non-functioning pseudogene, followed by targeted amplification of all exons within the active gene. This approach allows for 10-20x enrichment of CYP21A2 reads, and mutations detected are exclusively from the functional CYP21A2 locus.
NEXTFLEX® Congenital Adrenal Hyperplasia Amplicon PanelNEWBORN SYNDROMES & INFERTILITY
Gene Content CYP21A2 exons
Low Sample Requirement > 20 ng DNA from fresh or frozen samples
Coverage Uniformity 100% at 0.2x mean coverage
On-target Reads > 92%
High Sample IndexingUp to 384 samples with 100x coverage on a single Illumina® 2x150 MiSeq® lane for germline mutations
Efficient Workflow < 3 hrs hands-on time
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INNOVACIÓN TECNOLÓGICA PARA LABORATORIO
chr6 CYP21A2 (19 amplicons)
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8
32
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512
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Amplicons arranged by their chromosomal position
Figure 1. Performance of NEXTFLEX® Congenital Adrenal Hyperplasia Amplicon Panel Library. (Left) Gel image of amplicon products. (Right) Uniformity coverage across 19 amplicons.
Ordering Information
The NEXTFLEX® Congenital Adrenal Hyperplasia Amplicon Panel, along with our other Panel offerings, is a powerful tool enabling the accurate detection and differentiation of mutations for clinical research in a rapid and high-throughput manner using NGS.
References1. Falhammar, H. et al. 2015. Biochemical and Genetic Diagnosis of 21-hydroxylase Deficiency. Endocrine. 50(2):306-314.2. Concolino, P. et al. 2013. Genes, Pseudogenes and like Genes: The case of 21-hydroxylase in Italian population. Clin. Chim. Acta 424:85-89. doi:
10.1016/j.cca.2013.05.019.3. Xu, Z. et al. 2013. Comprehensive Mutation Analysis of the CYP21A2 Gene: An Efficient Multistep Approach to the Molecular Diagnosis of Congenital
Adrenal Hyperplasia. J. Mol. Diagn. 15(6): 745-753.4. Haider, S. et al. 2012. Structure-Phenotype Correlations of Human
CATALOG # PRODUCT NAME QUANTITY
NOVA-4244-01 NEXTFLEX® CAH Amplicon Panel for Illumina® Platforms (8 Barcodes) 8 rxns
NOVA-4244-02 NEXTFLEX® CAH Amplicon Panel for Illumina® Platforms (48 Barcodes) 48 rxns
NOVA-4244-03 NEXTFLEX® CAH Amplicon Panel for Illumina® Platforms (Barcodes 1-96) 96 rxns
NOVA-4244-04 NEXTFLEX® CAH Amplicon Panel for Illumina® Platforms (Barcodes 97-192) 96 rxns
NOVA-4244-05 NEXTFLEX® CAH Amplicon Panel for Illumina® Platforms (Barcodes 193-288) 96 rxns
NOVA-4244-06 NEXTFLEX® CAH Amplicon Panel for Illumina® Platforms (Barcodes 289-384) 96 rxns
20% Uniformity Threshold
*Reagents easily compatible with Thermo Fisher® Ion Torrent™ platforms. Please inquire for details.For research use only. Not for use in diagnostic procedures.
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